O-Linked β-N-Acetylglucosaminyltransferase Substrate Specificity Is Regulated by Myosin Phosphatase Targeting and Other Interacting Proteins

Abstract

O-GlcNAc-transferase (OGT) substrate specificity is regulated by transiently interacting proteins. To further examine the regulation of OGT, we have identified 27 putative OGT-interacting proteins through a yeast two-hybrid screen. Two of these proteins, Trak1 (OIP106) and O-GlcNAcase, have been shown previously to interact with and regulate OGT. We demonstrate here that MYPT1 and CARM1 also interact with and target OGT. MYPT1 and CARM1 are substrates of OGT in vitro and in vivo. MYPT1 and CARM1 also function to alter OGT substrate specificity in vitro. Furthermore depletion of MYPT1 in Neuro-2a neuroblastoma cells alters GlcNAcylation of several proteins under basal conditions, suggesting that MYPT1 regulates OGT substrate specificity in vivo. O-GlcNAc-transferase (OGT) substrate specificity is regulated by transiently interacting proteins. To further examine the regulation of OGT, we have identified 27 putative OGT-interacting proteins through a yeast two-hybrid screen. Two of these proteins, Trak1 (OIP106) and O-GlcNAcase, have been shown previously to interact with and regulate OGT. We demonstrate here that MYPT1 and CARM1 also interact with and target OGT. MYPT1 and CARM1 are substrates of OGT in vitro and in vivo. MYPT1 and CARM1 also function to alter OGT substrate specificity in vitro. Furthermore depletion of MYPT1 in Neuro-2a neuroblastoma cells alters GlcNAcylation of several proteins under basal conditions, suggesting that MYPT1 regulates OGT substrate specificity in vivo. In metazoans, nuclear and cytoplasmic proteins are post-translationally modified on serines and threonines with O-GlcNAc 3The abbreviations used are: O-GlcNAc, O-linked β-N-acetylglucosamine; O-GlcNAcase, O-linked β-N-acetylglucosaminidase (β-N-acetylhexosaminidase, EC 3.2.1.52); OGT, O-linked β-N-acetylglucosaminyltransferase (protein N-acetylglucosaminyltransferase, EC 2.4.1.94); MYPT1, myosin phosphatase targeting subunit 1; CARM1, coactivator-associated arginine methyltransferase 1; siRNA, small interfering RNA; GST, glutathione S-transferase; PP1β, protein phosphatase 1β; HA, hemagglutinin. 3The abbreviations used are: O-GlcNAc, O-linked β-N-acetylglucosamine; O-GlcNAcase, O-linked β-N-acetylglucosaminidase (β-N-acetylhexosaminidase, EC 3.2.1.52); OGT, O-linked β-N-acetylglucosaminyltransferase (protein N-acetylglucosaminyltransferase, EC 2.4.1.94); MYPT1, myosin phosphatase targeting subunit 1; CARM1, coactivator-associated arginine methyltransferase 1; siRNA, small interfering RNA; GST, glutathione S-transferase; PP1β, protein phosphatase 1β; HA, hemagglutinin. in response to different extracellular signals, altering their biochemical and functional properties (for a review, see Ref. 1Copeland R.J. Bullen J.W. Hart G.W. Am. J. Physiol. 2008; 295: E17-E28Crossref PubMed Scopus (189) Google Scholar). The addition of the monosaccharide moiety is catalyzed by O-GlcNAc-transferase (OGT) using UDP-GlcNAc as the nucleotide sugar donor (2Haltiwanger R.S. Blomberg M.A. Hart G.W. J. PubMed Google the of the O-GlcNAc is catalyzed by Hart G.W. J. PubMed Google protein GlcNAcylation is to a is is catalyzed by by the PubMed Scopus Google GlcNAcylation is catalyzed by the of a PubMed Scopus Google Hart G.W. PubMed Scopus Google Scholar). substrate is the of OGT substrate specificity is proteins have been identified R.J. Bullen J.W. Hart G.W. Am. J. Physiol. 2008; 295: E17-E28Crossref PubMed Scopus (189) Google the of OGT is to substrates proteins is OGT to specificity in vitro. In OGT is to a of UDP-GlcNAc and the of OGT substrates is with different UDP-GlcNAc Hart G.W. J. PubMed Scopus Google Scholar). a been that the substrate specificity of OGT in regulated by UDP-GlcNAc have been shown to to the of the J. PubMed Scopus Google J. PubMed Scopus Google PubMed Google Scholar). in protein in GlcNAcylation of proteins that are of in UDP-GlcNAc of OGT been with a of OGT, regulation J. PubMed Scopus Google 2008; PubMed Scopus Google Scholar). The of OGT a to that of J. PubMed Scopus Google 2008; PubMed Scopus Google suggesting that OGT interact with a of proteins through Blomberg M.A. Hart G.W. J. PubMed Scopus Google Hart G.W. J. PubMed Scopus Google of been shown that Trak1 as with the of OGT, OGT to Hart G.W. J. PubMed Scopus Google Scholar). have OGT to regulated by interacting proteins PubMed Scopus Google J. J. J. PubMed Scopus Google 2008; PubMed Scopus Google Scholar). we have that in Neuro-2a cells the GlcNAcylation of targeting by Hart G.W. J. 2008; PubMed Scopus Google Scholar). in of interacting with the of OGT, with the of OGT. of OGT also been shown to interact with to of OGT to the 2008; PubMed Scopus Google is that of OGT through a of OGT and substrate To further we to OGT-interacting proteins using a yeast two-hybrid screen. we OGT also we demonstrate that MYPT1 and CARM1 function to alter OGT substrate neuroblastoma cells in modified and in a with and the yeast OGT to by and the using the the MYPT1, as a and J. M.A. J. PubMed Scopus Google Scholar). The by as a of PubMed Scopus Google Scholar). by by OGT a of J. Am. PubMed Scopus Google and MYPT1 Neuro-2a cells with and using to the a of a of and and as previously J. Am. PubMed Scopus Google Scholar). using the and to the and as previously with M.A. J. PubMed Scopus Google Scholar). in with The and the a with a of to The and and to the with the with with The and cells with using on and Neuro-2a and as previously Hart G.W. J. 2008; PubMed Scopus Google OGT MYPT1 OGT as MYPT1 O-GlcNAc of of Hart G.W. J. PubMed Scopus Google and in in of in using by and using to the in the The yeast two-hybrid to the with the with the to the to the yeast by on (for to and to the the the that by of these is in of 27 putative OGT-interacting proteins identified by yeast two-hybrid a by and protein protein protein protein protein protein phosphatase targeting subunit PubMed Scopus Google protein arginine methyltransferase protein protein protein protein and protein Hart G.W. J. PubMed Scopus Google and in a OGT and as previously with Hart G.W. J. 2008; PubMed Scopus Google Scholar). of of GST, in of of of with and by The protein using with and to OGT to MYPT1 with and of the in the with GST, as previously with Hart G.W. J. 2008; PubMed Scopus Google Scholar). and using of a substrate The and by as previously Hart G.W. J. 2008; PubMed Scopus Google on the function in The in O-GlcNAc as and the the by in the The of the the with The in the of The to and on the by a and OGT-interacting a OGT proteins, we a yeast two-hybrid of a using OGT as we to 27 putative OGT In previously interacting OGT PubMed Scopus Google and Trak1 as Hart G.W. J. PubMed Scopus Google Scholar). is to that the proteins identified to a of functional proteins to and the of the yeast two-hybrid we several of the yeast of the to interact with OGT, the protein interact with OGT we also to as a of the the of the protein to the on the of the putative OGT interact with OGT in of proteins on yeast with OGT and the proteins to and OGT and a to of shown are of and CARM1 with putative OGT the yeast two-hybrid MYPT1 and CARM1, further MYPT1 is a targeting subunit of as (for a review, see Ref. 2008; PubMed Scopus Google CARM1 is a arginine methyltransferase (for a review, see Ref. J. PubMed Scopus Google Neuro-2a cells with HA, MYPT1, CARM1 and their to interact with OGT. MYPT1 and CARM1 to with OGT and and CARM1 interact with OGT in Neuro-2a Neuro-2a cells with and OGT, PP1β, and Neuro-2a cells with and OGT and OGT, MYPT1, using and OGT and shown are of we that MYPT1 with OGT in vivo. and we to MYPT1 with OGT and CARM1 MYPT1 and CARM1 interact with OGT, we to substrates of OGT. we Neuro-2a cells with HA, and their GlcNAcylation using MYPT1 and CARM1 are in and and CARM1 are substrates of OGT in and in vitro. Neuro-2a cells with and O-GlcNAc and Neuro-2a cells with and O-GlcNAc and and as substrates in the of The of the with in the and as substrates in the of The of the with in the shown are of that MYPT1 and CARM1 are OGT we in vitro OGT using proteins. MYPT1 and CARM1, GST, are substrates OGT in vitro and MYPT1 to a substrate CARM1 MYPT1 CARM1 with and CARM1 OGT in have that OGT is to substrates by interacting proteins Hart G.W. J. PubMed Scopus Google Hart G.W. J. PubMed Scopus Google PubMed Scopus Google J. J. J. PubMed Scopus Google 2008; PubMed Scopus Google Hart G.W. J. 2008; PubMed Scopus Google Scholar). we to MYPT1 CARM1 to the substrate of OGT. We OGT in the of to and of the of OGT substrates we to the proteins by the a MYPT1 The proteins to and as substrates OGT in the of are several proteins that to OGT substrates in the of suggesting that MYPT1 alter OGT substrate specificity in vitro. a substrate OGT in the of proteins, that are proteins that also the of OGT We also to see MYPT1 OGT myosin in vitro. MYPT1 on the GlcNAcylation of myosin in vitro and CARM1 alter OGT substrate in vitro. a MYPT1 of as substrate in the of and The of the with in the a CARM1 of as substrate in the of and The of the with in the shown are of we CARM1 function in in vitro OGT that are a proteins that are OGT substrates in the of that CARM1 alter OGT substrate specificity in vitro. also that CARM1 GlcNAcylation of OGT OGT in MYPT1 alters OGT substrate in we to the of of MYPT1 on protein depletion of MYPT1 in Neuro-2a cells on OGT, O-GlcNAcase, PP1β, protein GlcNAcylation In that depletion of MYPT1 the in GlcNAcylation of proteins the of OGT and suggesting that MYPT1 is GlcNAcylation of proteins under basal In of CARM1 to on protein GlcNAcylation under basal of OGT with CARM1 to CARM1 and in to proteins in response to different is the function of OGT as a substrate specificity is by using a of proteins to OGT to we the of putative OGT-interacting proteins and the of MYPT1 and CARM1 as OGT MYPT1 to a of OGT under basal conditions, CARM1 is to regulate OGT under is by interacting proteins under different to are on the under CARM1 regulates OGT. CARM1 is arginine methyltransferase substrates of the J. PubMed Scopus Google Scholar). nuclear are proteins CARM1 to the of to proteins to J. PubMed Scopus Google Scholar). is that under these CARM1 OGT to of the to OGT been shown to regulate the of to a of different to a GlcNAcylation in the of the of OGT by CARM1 to the of OGT and protein GlcNAcylation in the regulation of by nuclear are also to the proteins that are to OGT by MYPT1 and The of these substrates to the functional of MYPT1 and CARM1 in OGT. myosin are J. PubMed Scopus Google J. J. PubMed Scopus Google MYPT1 the of OGT MYPT1 and OGT are to interact with PubMed Scopus Google Hart G.W. J. PubMed Scopus Google MYPT1 to the of OGT with the function and of on OGT are 2008; PubMed Scopus Google is that MYPT1 also to OGT under been shown that MYPT1 is to and cells MYPT1 have to PubMed Scopus Google Scholar). In been shown that of MYPT1 in in in and cells with a in and by 2008; PubMed Scopus Google J. PubMed Scopus Google J. PubMed Scopus Google Scholar). We have shown that of O-GlcNAc by of OGT alters and also Hart G.W. J. PubMed Scopus Google Scholar). the of MYPT1 is also to protein GlcNAcylation is under we have shown here that MYPT1 regulates OGT basal conditions, is that MYPT1 also regulate OGT OGT to a different of that a protein of OGT to different of substrates on the is with of protein MYPT1 the of myosin PubMed Scopus Google the of 2008; PubMed Scopus Google Scholar). is to OGT by MYPT1, a the of the of OGT with in GlcNAcylation of different proteins and to protein and we a of OGT-interacting proteins. to see of these proteins functional as OGT to substrate In metazoans, nuclear and cytoplasmic proteins are post-translationally modified on serines and threonines with O-GlcNAc 3The abbreviations used are: O-GlcNAc, O-linked β-N-acetylglucosamine; O-GlcNAcase, O-linked β-N-acetylglucosaminidase (β-N-acetylhexosaminidase, EC 3.2.1.52); OGT, O-linked β-N-acetylglucosaminyltransferase (protein N-acetylglucosaminyltransferase, EC 2.4.1.94); MYPT1, myosin phosphatase targeting subunit 1; CARM1, coactivator-associated arginine methyltransferase 1; siRNA, small interfering RNA; GST, glutathione S-transferase; PP1β, protein phosphatase 1β; HA, hemagglutinin. 3The abbreviations used are: O-GlcNAc, O-linked β-N-acetylglucosamine; O-GlcNAcase, O-linked β-N-acetylglucosaminidase (β-N-acetylhexosaminidase, EC 3.2.1.52); OGT, O-linked β-N-acetylglucosaminyltransferase (protein N-acetylglucosaminyltransferase, EC 2.4.1.94); MYPT1, myosin phosphatase targeting subunit 1; CARM1, coactivator-associated arginine methyltransferase 1; siRNA, small interfering RNA; GST, glutathione S-transferase; PP1β, protein phosphatase 1β; HA, hemagglutinin. in response to different extracellular signals, altering their biochemical and functional properties (for a review, see Ref. 1Copeland R.J. Bullen J.W. Hart G.W. Am. J. Physiol. 2008; 295: E17-E28Crossref PubMed Scopus (189) Google Scholar). The addition of the monosaccharide moiety is catalyzed by O-GlcNAc-transferase (OGT) using UDP-GlcNAc as the nucleotide sugar donor (2Haltiwanger R.S. Blomberg M.A. Hart G.W. J. PubMed Google the of the O-GlcNAc is catalyzed by Hart G.W. J. PubMed Google Scholar). In protein GlcNAcylation is to a is is catalyzed by by the PubMed Scopus Google GlcNAcylation is catalyzed by the of a PubMed Scopus Google Hart G.W. PubMed Scopus Google Scholar). substrate is the of OGT substrate specificity is proteins have been identified R.J. Bullen J.W. Hart G.W. Am. J. Physiol. 2008; 295: E17-E28Crossref PubMed Scopus (189) Google the of OGT is to substrates proteins is OGT to specificity in vitro. In OGT is to a of UDP-GlcNAc and the of OGT substrates is with different UDP-GlcNAc Hart G.W. J. PubMed Scopus Google Scholar). a been that the substrate specificity of OGT in regulated by UDP-GlcNAc have been shown to to the of the J. PubMed Scopus Google J. PubMed Scopus Google PubMed Google Scholar). in protein in GlcNAcylation of proteins that are of in UDP-GlcNAc The of OGT been with a of OGT, regulation J. PubMed Scopus Google 2008; PubMed Scopus Google Scholar). The of OGT a to that of J. PubMed Scopus Google 2008; PubMed Scopus Google suggesting that OGT interact with a of proteins through Blomberg M.A. Hart G.W. J. PubMed Scopus Google Hart G.W. J. PubMed Scopus Google Scholar). In of been shown that Trak1 as with the of OGT, OGT to Hart G.W. J. PubMed Scopus Google Scholar). have OGT to regulated by interacting proteins PubMed Scopus Google J. J. J. PubMed Scopus Google 2008; PubMed Scopus Google Scholar). we have that in Neuro-2a cells the GlcNAcylation of targeting by Hart G.W. J. 2008; PubMed Scopus Google Scholar). in of interacting with the of OGT, with the of OGT. of OGT also been shown to interact with to of OGT to the 2008; PubMed Scopus Google Scholar). is that of OGT through a of OGT and substrate To further we to OGT-interacting proteins using a yeast two-hybrid screen. we OGT also we demonstrate that MYPT1 and CARM1 function to alter OGT substrate neuroblastoma cells in modified and in a with and the yeast OGT to by and the using the the MYPT1, as a and J. M.A. J. PubMed Scopus Google Scholar). The by as a of PubMed Scopus Google Scholar). by by OGT a of J. Am. PubMed Scopus Google and MYPT1 Neuro-2a cells with and using to the a of a of and and as previously J. Am. PubMed Scopus Google Scholar). using the and to the and as previously with M.A. J. PubMed Scopus Google Scholar). in with The and the a with a of to The and and to the with the with with The and cells with using on and Neuro-2a and as previously Hart G.W. J. 2008; PubMed Scopus Google OGT MYPT1 OGT as MYPT1 O-GlcNAc of of Hart G.W. J. PubMed Scopus Google and in in of in using by and using to the in the The yeast two-hybrid to the with the with the to the to the yeast by on (for to and to the the the that by of these is in of 27 putative OGT-interacting proteins identified by yeast two-hybrid a by and protein protein protein protein protein protein phosphatase targeting subunit PubMed Scopus Google protein arginine methyltransferase protein protein protein protein and protein Hart G.W. J. PubMed Scopus Google and in a OGT and as previously with Hart G.W. J. 2008; PubMed Scopus Google Scholar). of of GST, in of of of with and by The protein using with and to OGT to MYPT1 with and of the in the with GST, as previously with Hart G.W. J. 2008; PubMed Scopus Google Scholar). and using of a substrate The and by as previously Hart G.W. J. 2008; PubMed Scopus Google on the function in The in O-GlcNAc as and the the by in the The of the the with The in the of The to and on the by a and neuroblastoma cells in modified and in a with and the yeast OGT to by and the using the the MYPT1, as a and J. M.A. J. PubMed Scopus Google Scholar). The by as a of PubMed Scopus Google Scholar). by by The OGT a of J. Am. PubMed Scopus Google Scholar). and MYPT1 Neuro-2a cells with and using to the a of a of and and as previously J. Am. PubMed Scopus Google Scholar). using the and to the and as previously with M.A. J. PubMed Scopus Google Scholar). in with The and the a with a of to The and and to the with the with with The and cells with using on and Neuro-2a and as previously Hart G.W. J. 2008; PubMed Scopus Google Scholar). OGT MYPT1 OGT as MYPT1 O-GlcNAc of of Hart G.W. J. PubMed Scopus Google and in in of in using by and using to the in the The yeast two-hybrid to the The with the with the to the to the yeast by on (for to and to the the the that by of these is in OGT and as previously with Hart G.W. J. 2008; PubMed Scopus Google Scholar). of of GST, in of of of with and by The protein using with and to OGT to MYPT1 with and of the in the with GST, OGT as previously with Hart G.W. J. 2008; PubMed Scopus Google Scholar). and using of a substrate The and by as previously Hart G.W. J. 2008; PubMed Scopus Google Scholar). on the function in The in O-GlcNAc as and the the by in the The of the the with The in the of The to and on the by a and OGT-interacting a OGT proteins, we a yeast two-hybrid of a using OGT as we to 27 putative OGT In previously interacting OGT PubMed Scopus Google and Trak1 as Hart G.W. J. PubMed Scopus Google Scholar). is to that the proteins identified to a of functional proteins to and the of the yeast two-hybrid we several of the yeast of the to interact with OGT, the protein interact with OGT we also to as a of the the of the protein to the on the of the and CARM1 with putative OGT the yeast two-hybrid MYPT1 and CARM1, further MYPT1 is a targeting subunit of as (for a review, see Ref. 2008; PubMed Scopus Google CARM1 is a arginine methyltransferase (for a review, see Ref. J. PubMed Scopus Google Neuro-2a cells with HA, MYPT1, CARM1 and their to interact with OGT. MYPT1 and CARM1 to with OGT and and CARM1 interact with OGT in Neuro-2a Neuro-2a cells with and OGT, PP1β, and Neuro-2a cells with and OGT and OGT, MYPT1, using and OGT and shown are of we that MYPT1 with OGT in vivo. and we to MYPT1 with OGT and CARM1 MYPT1 and CARM1 interact with OGT, we to substrates of OGT. we Neuro-2a cells with HA, and their GlcNAcylation using MYPT1 and CARM1 are in and and CARM1 are substrates of OGT in and in vitro. Neuro-2a cells with and O-GlcNAc and Neuro-2a cells with and O-GlcNAc and and as substrates in the of The of the with in the and as substrates in the of The of the with in the shown are of that MYPT1 and CARM1 are OGT we in vitro OGT using proteins. MYPT1 and CARM1, GST, are substrates OGT in vitro and MYPT1 to a substrate CARM1 MYPT1 CARM1 with and CARM1 OGT in have that OGT is to substrates by interacting proteins Hart G.W. J. PubMed Scopus Google Hart G.W. J. PubMed Scopus Google PubMed Scopus Google J. J. J. PubMed Scopus Google 2008; PubMed Scopus Google Hart G.W. J. 2008; PubMed Scopus Google Scholar). we to MYPT1 CARM1 to the substrate of OGT. We OGT in the of to and of the of OGT substrates we to the proteins by the a MYPT1 The proteins to and as substrates OGT in the of are several proteins that to OGT substrates in the of suggesting that MYPT1 alter OGT substrate specificity in vitro. a substrate OGT in the of proteins, that are proteins that also the of OGT We also to see MYPT1 OGT myosin in vitro. MYPT1 on the GlcNAcylation of myosin in vitro and CARM1 alter OGT substrate in vitro. a MYPT1 of as substrate in the of and The of the with in the a CARM1 of as substrate in the of and The of the with in the shown are of we CARM1 function in in vitro OGT that are a proteins that are OGT substrates in the of that CARM1 alter OGT substrate specificity in vitro. also that CARM1 GlcNAcylation of OGT OGT in MYPT1 alters OGT substrate in we to the of of MYPT1 on protein depletion of MYPT1 in Neuro-2a cells on OGT, O-GlcNAcase, PP1β, protein GlcNAcylation In that depletion of MYPT1 the in GlcNAcylation of proteins the of OGT and suggesting that MYPT1 is GlcNAcylation of proteins under basal In of CARM1 to on protein GlcNAcylation under basal of OGT with CARM1 to CARM1 and in OGT-interacting a OGT proteins, we a yeast two-hybrid of a using OGT as we to 27 putative OGT In previously interacting OGT PubMed Scopus Google and Trak1 as Hart G.W. J. PubMed Scopus Google Scholar). is to that the proteins identified to a of functional proteins to and To the of the yeast two-hybrid we several of the yeast of the to interact with OGT, the protein interact with OGT we also to as a of the the of the protein to the on the of the MYPT1 and CARM1 with putative OGT the yeast two-hybrid MYPT1 and CARM1, further MYPT1 is a targeting subunit of as (for a review, see Ref. 2008; PubMed Scopus Google CARM1 is a arginine methyltransferase (for a review, see Ref. J. PubMed Scopus Google Scholar). We Neuro-2a cells with HA, MYPT1, CARM1 and their to interact with OGT. MYPT1 and CARM1 to with OGT and we that MYPT1 with OGT in vivo. and we to MYPT1 with OGT MYPT1 and CARM1 MYPT1 and CARM1 interact with OGT, we to substrates of OGT. we Neuro-2a cells with HA, and their GlcNAcylation using MYPT1 and CARM1 are in and To that MYPT1 and CARM1 are OGT we in vitro OGT using proteins. MYPT1 and CARM1, GST, are substrates OGT in vitro and MYPT1 to a substrate CARM1 MYPT1 CARM1 with OGT. MYPT1 and CARM1 OGT in have that OGT is to substrates by interacting proteins Hart G.W. J. PubMed Scopus Google Hart G.W. J. PubMed Scopus Google PubMed Scopus Google J. J. J. PubMed Scopus Google 2008; PubMed Scopus Google Hart G.W. J. 2008; PubMed Scopus Google Scholar). we to MYPT1 CARM1 to the substrate of OGT. We OGT in the of to and of the of OGT substrates we to the proteins by the a MYPT1 The proteins to and as substrates OGT in the of are several proteins that to OGT substrates in the of suggesting that MYPT1 alter OGT substrate specificity in vitro. a substrate OGT in the of proteins, that are proteins that also the of OGT We also to see MYPT1 OGT myosin in vitro. MYPT1 on the GlcNAcylation of myosin in vitro we CARM1 function in in vitro OGT that are a proteins that are OGT substrates in the of that CARM1 alter OGT substrate specificity in vitro. also that CARM1 GlcNAcylation of OGT MYPT1 OGT in MYPT1 alters OGT substrate in we to the of of MYPT1 on protein depletion of MYPT1 in Neuro-2a cells on OGT, O-GlcNAcase, PP1β, protein GlcNAcylation In that depletion of MYPT1 the in GlcNAcylation of proteins the of OGT and suggesting that MYPT1 is GlcNAcylation of proteins under basal In of CARM1 to on protein GlcNAcylation under basal of OGT with CARM1 to CARM1 and in to proteins in response to different is the function of OGT as a substrate specificity is by using a of proteins to OGT to we the of putative OGT-interacting proteins and the of MYPT1 and CARM1 as OGT MYPT1 to a of OGT under basal conditions, CARM1 is to regulate OGT under are on the under CARM1 regulates OGT. CARM1 is arginine methyltransferase substrates of the J. PubMed Scopus Google Scholar). nuclear are proteins CARM1 to the of to proteins to J. PubMed Scopus Google Scholar). is that under these CARM1 OGT to of the to OGT been shown to regulate the of to a of different to a GlcNAcylation in the of the of OGT by CARM1 to the of OGT and protein GlcNAcylation in the regulation of by nuclear are also to the proteins that are to OGT by MYPT1 and The of these substrates to the functional of MYPT1 and CARM1 in OGT. myosin are J. PubMed Scopus Google J. J. PubMed Scopus Google MYPT1 the of OGT MYPT1 and OGT are to interact with PubMed Scopus Google Hart G.W. J. PubMed Scopus Google MYPT1 to the of OGT with the function and of on OGT are 2008; PubMed Scopus Google is that MYPT1 also to OGT under been shown that MYPT1 is to and cells MYPT1 have to PubMed Scopus Google Scholar). In been shown that of MYPT1 in in in and cells with a in and by 2008; PubMed Scopus Google J. PubMed Scopus Google J. PubMed Scopus Google Scholar). 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PubMed Scopus Google Scholar). nuclear are proteins CARM1 to the of to proteins to J. PubMed Scopus Google Scholar). is that under these CARM1 OGT to of the to OGT been shown to regulate the of to a of different to a GlcNAcylation in the of the of OGT by CARM1 to the of OGT and protein GlcNAcylation in the regulation of by nuclear We are also to the proteins that are to OGT by MYPT1 and The of these substrates to the functional of MYPT1 and CARM1 in OGT. myosin are J. PubMed Scopus Google J. J. PubMed Scopus Google MYPT1 the of OGT MYPT1 and OGT are to interact with PubMed Scopus Google Hart G.W. J. PubMed Scopus Google MYPT1 to the of OGT with the function and of on OGT are 2008; PubMed Scopus Google is that MYPT1 also to OGT under been shown that MYPT1 is to and cells MYPT1 have to PubMed Scopus Google Scholar). In been shown that of MYPT1 in in in and cells with a in and by 2008; PubMed Scopus Google J. PubMed Scopus Google J. PubMed Scopus Google Scholar). 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